Introduction to experimental biophysics : biological methods for physical scientists /: biological methods for physical scientists. ([2012])

• Record Type: Book
• Other Titles: Experimental biophysics
• Further Information: Note: Jay Nadeau.
• Other Names: Nadeau, Jay L
• Contents: Introduction to the background. Basic biochemistry ; Energies and potentials ; Principles of spectroscopy ; Cells ; DNA, RNA, replication, and transcription ; Translation and the genetic code ; Protein folding and trafficking ; Alternative genetics ; What is cloning? ; Design of a molecular biology experiment and how to use this book -- Molecular cloning of DNA and RNA. Obtaining and storing plasmids ; Selection of an appropriate E. coli amplification strain : transformation of E. coli with plasmid ; Plasmid amplification and purification ; Plasmid restriction mapping and agarose gel electrophoresis ; An example of cloning experiment ; Cloning by the polymerase chain reaction ; Sequencing ; RNA methods ; Southern and northern blots ; Solutions and large cloning problems and multiple inserts ; Mutagenesis and directed evolution ; Microarrays ; Summary and hints -- Expression of genes in bacteria, yeast, and cultured mammalian cells. Expressing genes in microorganisms ; Mammalian cell culture ; Transfection of mammalian cells I : standard techniques ; Transfection of mammalian cells II : specialized physical methods for special occasions ; Transfection of mammalian cells III : viruses ; Summary. Protein expression methods. Expression systems ; Identification of a DNA source ; Selecting an expression vector ; Sub-cloning into an expression vector ; Selection of an expression strain or cell line ; Protein expression ; SDS-PAGE ; Protein isolation and purification ; Chromatography ; Buffer exchange and concentration ; Example experiment : expression and purification of fluorescent protein dronpa ; Conclusions and final remarks -- Protein crystallization. Crystallization of macromolecules ; Preparation of proteins for crystallization ; Components of crystallization solutions ; Other factors affecting crystallization ; Crystallization strategies ; Example experiment : lysozyme ; Data collection and structure determination using x-ray crystallography ; A special case : membrane proteins ; Troubleshooting Q & A ; Conclusions and final remarks -- Introduction to biological light microscopy. The physics of microscopy : magnification and resolution ; Anatomy of a biological microscope ; Brightfield imaging techniques ; Basic fluorescence microscopy ; Fluorophores for cell labeling ; Fluorescent proteins ; Multispectral imaging using acousto-optical tunable filters ; Advanced techniques ; Summary and remarks. Quantitative cell culture technique. Quantifying bacterial growth and death ; Quantifying mammalian cells ; Flow cytometry ; Example experiment : determining Leukemic B-cells and T-cells by flow cytometry ; Quantifying viruses ; Measuring cell populations using quantitative PCR ; Summary and final remarks -- Semiconductor nanoparticles (quantum dots). Quantum dot properties and synthesis ; QD applications ; Example experiment : conjugation of quantum dots to Dopamine and quantifying the effects on Fluorescence per molecule bound ; Summary and remarks -- Gold nanoparticles. The physics of scattering and spherical metal nanoparticles ; Synthesis of gold nanoparticles ; Characterization and surface modification of gold nanoparticles ; Applications for colorimetric detection and microscopy ; Sample experiment : labeling cells with Lectin-Tagged Au Nanoparticles ; Applications in surface-enhanced raman scattering ; Gold nanoparticles as photothermal transducers ; Conclusion. Surface functionalization techniques. Preparing monolayers using functional silanes or thiols ; Techniques for characterizing surface monolayers ; Functionalization of modified surfaces using cross-linkers ; Example experiment : preparing a Silane-Biotin-Streptavidin sandwich on SiO₂ features on a Si Chip ; Preventing nonspecific binding of biomolecules ; Assembling membrane proteins on surfaces ; Testing the function of immobilized proteins ; Conclusion and final remarks -- Electrophysiology. Physical basis and circuit models ; Solutions and blockers ; Instrumentation ; Lipid bilayer setup ; Cell patch-clamp setup : What is needed? ; The art and magic of pipette pulling ; Step-by-step guide to performing a whole-cell recording ; Example experiment : whole-cell recording on cells transfected with K+ channels and GFP ; A brief introduction to single-channel modeling and data analysis ; Network recording ; Conclusions and final remarks -- Spectroscopy tools and techniques. Guiding principles ; UV-Vis absorbance spectroscopy ; Fluorescence spectroscopy ; Time-resolved emission ; Time-resolved absorption ; Infrared spectroscopy ; Nuclear magnetic resonance ; Electron paramagnetic resonance spectroscopy ; X-ray spectroscopy ; Example experiment : characterization of CdSe/ZnS nanoparticle bioconjugate using UV-Vis, Fluorescence emission, time-resolved emission, FTIR, and EPR spectroscopy ; Final comments -- Appendix. Common solutions ; Microbial growth media ; Agarose gel recipes ; Protein gel recipes ; Restriction endonucleases ; Common nucleic acide modifying enzymes ; Fluorescent dyes and quenchers ; Fluorescent proteins.
• Publisher Details: Boca Raton, Fla : CRC Press
• Publication Date: 2012
• Copyright Date: 2012
• Extent: 1 online resource, illustrations (some color)
• Subjects: 571.4/0724; Biophysics -- Experiments -- Technique; Biophysics -- methods; Biochemistry -- methods; Molecular Biology -- methods; Biophysik; Experimentalphysik; Biofysik
• Languages: English
• ISBNs: 9781439897409; 1439897409
• Related ISBNs: 9781439829530; 1439829535
• Notes: Note: Includes bibliographical references and index.; Note: Print version record.
• Access Rights: Legal Deposit; Only available on premises controlled by the deposit library and to one user at any one time; The Legal Deposit Libraries (Non-Print Works) Regulations (UK).
• Access Usage: Restricted: Printing from this resource is governed by The Legal Deposit Libraries (Non-Print Works) Regulations (UK) and UK copyright law currently in force.